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Related Concept Videos

Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Phosphorylation01:02

Phosphorylation

The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...

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Updated: May 29, 2026

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
10:17

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

Published on: April 29, 2022

Phosphoprotein stability in clinical tissue and its relevance for reverse phase protein microarray technology.

Virginia Espina1, Claudius Mueller, Lance A Liotta

  • 1Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA. vespina@gmu.edu

Methods in Molecular Biology (Clifton, N.J.)
|September 9, 2011
PubMed
Summary

Proper tissue handling is crucial for accurate cell signaling analysis. Immediate preservation of phosphoproteins post-procurement ensures reliable measurement of kinase pathway activity in tumors.

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Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis
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Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis

Published on: April 1, 2022

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Last Updated: May 29, 2026

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
10:17

A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors

Published on: April 29, 2022

Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis
07:26

Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis

Published on: April 1, 2022

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cancer Research

Background:

  • Phosphorylated proteins indicate cell signaling activity within kinase networks.
  • Cell signaling pathway status reflects in vivo cellular and tissue activity.
  • Tumor growth can be linked to specific phosphorylated proteins and pathways.

Purpose of the Study:

  • To investigate factors affecting phosphoprotein stability in tissue after procurement.
  • To establish tissue procurement guidelines for accurate phosphoprotein quantification.
  • To minimize pre-analytical variability in molecular analysis.

Main Methods:

  • Utilized Reverse Phase Protein Microarrays (RPMAs) for quantitative measurement of phosphoproteins.
  • Analyzed hundreds of phosphorylated signal proteins from small tissue samples.
  • Studied the influence of ex vivo delay and tissue handling on phosphoprotein levels.

Main Results:

  • Pre-analytical variability significantly impacts downstream molecular analysis.
  • Ex vivo tissue processing can alter phosphoprotein levels, not reflecting in vivo states.
  • Identified critical factors influencing phosphoprotein stability post-procurement.

Conclusions:

  • Immediate stabilization of phosphoproteins after tissue procurement is essential.
  • Established tissue procurement guidelines for clinical research focused on RPMA phosphoprotein quantification.
  • Accurate assessment of kinase networks requires preserving phosphoprotein integrity.